Incorporation of cytochrome oxidase into cardiolipin bilayers and induction of nonlamellar phases.

Cytochrome oxidase from beef heart has been lipid-substituted with beef heart cardiolipin. The lipid phase behavior and protein aggregation state of the reconstituted complexes have been studied with 31P NMR, freeze-fracture electron microscopy, and saturation-transfer ESR of the spin-labeled protein. In the absence of salt, the lipid has a lamellar arrangement, and the protein is integrated and uniformly distributed in the membrane vesicles and undergoes rapid rotational diffusion. The presence of the protein stabilizes the cardiolipin lamellar phase against salt-induced transitions to the inverted hexagonal phase. The threshold salt concentration becomes higher and the extent of conversion becomes lower with decreasing lipid:protein ratio. In high salt, lamellar-phase lipid with integrated protein coexists with hexagonal-phase lipid free of protein, and the rotational diffusion of the protein is drastically reduced as a result of the high packing density

Lipid-protein interactions in stacked and destacked thylakoid membranes and the influence of phosphorylation and illumination. Spin label ESR studies.

The effects of membrane destacking, protein phosphorylation, and continuous illumination have been studied in pea thylakoid membranes using ESR spectroscopy of an incorporated spin-labelled phosphatidylglycerol. This spin-labelled analogue of an endogenous thylakoid lipid has previously been shown to exhibit a selectivity of interaction with thylakoid proteins. Neither destacking, phosphorylation nor illumination was found to change the ESR spectra appreciably, suggesting that for phosphatidylglycerol at least, neither the number of protein-associated membrane lipids nor their pattern of selectivity was altered. The redistribution of the thylakoid protein complexes in the membrane, under these various conditions, therefore takes place with conservation of the properties of the lipid/protein interface

Solution to the Equations of the Moment Expansions

We develop a formula for matching a Taylor series about the origin and an asymptotic exponential expansion for large values of the coordinate. We test it on the expansion of the generating functions for the moments and connected moments of the Hamiltonian operator. In the former case the formula produces the energies and overlaps for the Rayleigh-Ritz method in the Krylov space. We choose the harmonic oscillator and a strongly anharmonic oscillator as illustrative examples for numerical test. Our results reveal some features of the connected-moments expansion that were overlooked in earlier studies and applications of the approach

Nitrogenase activity associated with codium species from New Zealand marine habitats

Nitrogenase activity, measured as acetylene reduction, was recorded at rates up to 1028 nmol.h \g * dry weight for Codium adhaerens (Cabr.) Ag. var. convolutum Dellow and Codium fragile (Sur.) Hariot subsp. tomentosoides (Van Goor) Silva collected from New Zealand habitats. In both species the ability to reduce acetylene is invariably associated with the presence of a heterocystous blue-green alga, Calothrix sp., epiphytic or embedded in the Codium thallus. A highly significant (P < 0.001) correlation between heterocyst frequency and nitrogenase activity was found. Nitrogenase and net photosynthesis of the Codium-Calothrix system have different steady-state responses to light intensity, and the kinetics of the two processes also differ in that nitrogenase is slow to respond to illumination or darkening. Glucose additions to Codium did not significantly increase nitrogenase activity. Nitrogenase is relatively insensitive to oxygen tension over the range 0-1.0 atm (0-1.033 kgf.cnT2) and still occurs at 1.5 atm (1.55 kgf.cm"2); this condition is unique in all nitrogenase systems thus far reported. Collectively these facts suggest that Calothrix is the agent primarily responsible for nitrogenase activity in these Codium species

Consistent treatment of hydrophobicity in protein lattice models accounts for cold denaturation

The hydrophobic effect stabilizes the native structure of proteins by minimizing the unfavourable interactions between hydrophobic residues and water through the formation of a hydrophobic core. Here we include the entropic and enthalpic contributions of the hydrophobic effect explicitly in an implicit solvent model. This allows us to capture two important effects: a length-scale dependence and a temperature dependence for the solvation of a hydrophobic particle. This consistent treatment of the hydrophobic effect explains cold denaturation and heat capacity measurements of solvated proteins.Comment: Added and corrected references for design procedure in main text (p. 2) and in Supplemental Information (p. 8

Compound-specific amino acid ¹⁵N stable isotope probing of nitrogen assimilation by the soil microbial biomass using gas chromatography/combustion/isotope ratio mass spectrometry

RATIONALE: Organic nitrogen (N) greatly exceeds inorganic N in soils, but the complexity and heterogeneity of this important soil N pool make investigations into the fate of N‐containing additions and soil organic N cycling challenging. This paper details a novel approach to investigate the fate of applied N in soils, generating quantitative measures of microbial assimilation and of newly synthesized soil protein. METHODS: Laboratory incubation experiments applying (15)N‐ammonium, (15)N‐nitrate and (15)N‐glutamate were carried out and the high sensitivity and selectivity of gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS) exploited for compound‐specific (15)N stable isotope probing ((15)N‐SIP) of extracted incubation soil amino acids (AAs; as N‐acetyl, O‐isopropyl derivatives). We then describe the interpretation of these data to obtain a measure of the assimilation of the applied (15)N‐labelled substrate by the soil microbial biomass and an estimate of newly synthesised soil protein. RESULTS: The cycling of agriculturally relevant N additions is undetectable via bulk soil N content and δ (15)N values and AA concentrations. The assimilation pathways of the three substrates were revealed via patterns in AA δ (15)N values with time, reflecting known biosynthetic pathways (e.g. ammonium uptake occurs first via glutamate) and these data were used to expose differences in the rates and fluxes of the applied N substrates into the soil protein pool (glutamate > ammonium > nitrate). CONCLUSIONS: Our compound‐specific (15)N‐SIP approach using GC/C/IRMS offers a number of insights, inaccessible via existing techniques, into the fate of applied (15)N in soils and is potentially widely applicable to the study of N cycling in any soil, or indeed, in any complex ecosystem. © 2016 The Authors. Rapid Communications in Mass Spectrometry Published by John Wiley & Sons Ltd

Dry-mass sensing for microfluidics

We present an approach for interfacing an electromechanical sensor with a microfluidic device for the accurate quantification of the dry mass of analytes within microchannels. We show that depositing solutes onto the active surface of a quartz crystal microbalance by means of an on-chip microfluidic spray nozzle and subsequent solvent removal provides the basis for the real-time determination of dry solute mass. Moreover, this detection scheme does not suffer from the decrease in the sensor’s quality factor and the viscous drag present if the measurement is performed in a liquid environment, yet allows solutions to be analysed. We demonstrate the sensitivity and reliability of our approach by controlled deposition of nanogram levels of salt and protein from a micrometer-sized channel.We thank Alexander K. Buell, Igor Efimov, and Victor Ostanin for valuable discussions on QCM sensors and gratefully acknowledge financial support from the Swiss National Science Foundation (SNF), the Engineering and Physical Sciences Research Council (EPSRC), the Biotechnology and Biological Sciences Research Council (BBSRC), the European Research Council (ERC), as well as the Frances and Augustus Newman Foundation.Permission is granted to quote from an AIP publication with the customary acknowledgment of the source. Republication of an article or portions thereof (e.g., excerpts of greater than 400 words, figures, tables, etc.) in original form or in translation, as well as other types of reuse (such as use in course packs or electronic reserves) require formal permission from AIP and may be subject to fees. Although it is not a legal requirement for permission, as a courtesy, an author of the original article should be informed of any request for republication/reuse

Collaboration and the Generation of New Knowledge in Networked Innovation Systems: A Bibliometric Analysis

AbstractCanola, a high-value, export-oriented agricultural commodity, was developed in Canada over the course of 40 years in public institutions, driven by imported technology and imported research scientists. The evolution of canola R&D closely mirrors the evolution of the Triple Helix Models of innovation. Through the application of longitudinal citation analysis, using five-year intervals, publications from Canadian public institutions involved in canola R&D have been analyzed. In the most recent five- year interval, the relative citation rates of public sector research increased by 60% compared to the global average. A unique fixed-effect negative binomial regression model is used to demonstrate the critical relationship between the institutional arrangement that governs collaboration and the production of knowledge that underscores technological innovation